• 한국식품영양학회지
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• 제10권2호
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• pp.151-159
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• 1997

근원섬유단백질의 이온강도에 따른 Ca-ATPase 활성, Mg-ATPase 활성 및 EDTA-ATPase 활성은 오징어와 대합에서 그 차이점이 뚜렷하였으며, activity-pH curve에서 오징어 actomyosin의 Ca-ATPase 활성은 biphasic response가 소실되었고 대합의 actomyosin은 미약한 bipasic response가 나타났다. 또한 저농도의 dioxane에 의하여 오징어의 근원섬유단백질은 급격한 활성의 감소를 보였으나 대합의 근원섬유단백질은 활성이 증가되었다. 그리고 에탄올과 메탄올은 오징어와 대합의 myosin 및 MM에 대하여 저농도에서 활성을 증가시켰다. 한편 NEM으로 근원섬유단백질을 modification시키며 10-6M 이하의 NEM 농도에서는 활성이 증가되었으나 10-5M 이상의 농도가 되면 활성의 급격한 감소가 나타났다.

• 대한한의학회지
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• 제17권2호
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• pp.264-276
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• 1996

This study was carried out to evaluate the effect of Samhwasan on the Na-K-ATPase activity of heart muscle. The Na-K-ATPase activity was prepared from rabbit heart ventricles. Samhwasan markedly inhibited the Na- K - ATPase activity in a dose-dependent manner with an estimated $I_{50}$ of 0.56%. Hill coefficient was 1.70, indicating that the enzyme has more than one binding site for the Samhwasan. Inhibition of enzyme activity by Samhwasan increased as pretreatment time was prolonged. Inhibition by the drug was not affected by a change in enzyme protein concentration. Kinetic studies of substrate activation of the enzyme indicated classical noncompetitive inhibition, showing significant reduction in Vmax without a change in Km value. Inhibitory effect by Samhwasan was not altered by changes in concentration of $Mg^{2+}$, $Na^+$ or $K^+$, dithiothreitol. a sulfhydryl reducing reagent, did not protect the inhibition of Na-K-ATPase activity by Samhwasan combination of Samhwasan and ouabain showed a cumulative inhibition fashion. These results suggest that Samhwasan inhibits Na-K-ATPase activity of heart ventricles with an unique binding site different from that of ATP, $Mg^{2+}$, $Na^+$ or $K^+$ and ouabain.

• 한국동물학회지
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• 제20권2호
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• pp.101-107
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• 1977

토끼의 골격근 小胞體의 ATPase 活性과 Ca 輸送에 대한 sodium azide, cAMP, G-strophanthin 및 dicumarol의 영향을 측정하였다. Sodium azide(0.05mM)와 G-strophanthin(0.25nM)은 ATPase活性과 Ca 輸送能에 아무런 영향도 미치지 아니하였다. cAMP$(1 \\times 10^-6 M \\sim 5 \\times 10^-4 M)$는 ATPase 活性에는 아무런 영향도 미치지 않았으나 Ca 輸送은 억제하였다. Dicumarol(0.05mM)도 ATPase 活性에는 영향이 없었으나 小胞體의 $8,000 \\sim 12,000 \\times G$分劃에서의 Ca 輸送을 억제하였다.

• 한국미생물·생명공학회지
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• 제21권1호
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• pp.30-35
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• 1993

E.coli에서 발견된 ATP-dependent 효소인 Clp효소 중에서 Clp A의 ATPase 활성에 대한 영향을 검토하였다. Clp효소의 limiting amount으로 나타난 specific 활성은 일정하게 증가하는 효소의존성을 보였다. ATPase 활성을 나타내고 있는 ClP A는 casein에 의하여 활성화되어지며 2분자의 ATP가 결합하고 ATPase 활성을 나타내기 위한 ATP의 분해는 Clp효소의 단백질 분해 활성에 필요하다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권3호
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• pp.192-198
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• 2003

BiP, immunoglobulin binding protein, is an ER homologue of Hsp70. However, unlit other Hsp70 proteins, regulatory protein(s) for BiP has not been identified. Here, we demo strafed the presence of potential regulatory proteins for BiP using a pull -down assay. Since BiP can bind any unfolded protein, only the ATPase domain of BiP was used for the pull -down assay in order to minimize nonspecific binding. The ATPase domain was cloned to produce recombinant protein, which was then conjugated to CNBr-activated agarose. The structural conformation and ATP hydrolysis activity of the recombinant ATPase domain were similar to those of the native protein, light proteins from metabolically labeled mouse plasmacytoma cells specifically bound to the recombinant ATPase protein. The binding of these proteins was inhibited by excess amounts of free ATPase protein, and was dependent on the presence of ATP. These proteins were eluted by ADP. Of these proteins, Grp170 and BiP where identified. while the other were not identified as known ER proteins, from Western blot analyses. The presence of the ATPase-binding proteins for BiP was first demonstrated in this study, and our data suggest similar regulatory machinery for BiP may exist in the ER, as found in prokaryotes and other cellular compartments.

• 한국잠사곤충학회지
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• 제42권1호
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• pp.14-23
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• 2000

The mitochondrial genome of domesticated silkworm (Bombyx mori) was mapped with five restriction endonucleases (BamHI, EcoRI, HindIII, PstI and XbaI), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5$^1$and 3$^1$region for full genome set of eleven mitochondrial clones, the seven mitochondrial genes (NADH dehydrogenase 6, ATPase 6, ATPase 8, tRN $A^{Lys}$, tRN $A^{Asp}$, tRN $A^{Thr}$ and tRN $A^{Phe}$ of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRN $A^{Lys}$, tRN $A^{Asp}$ and tRN $A^{Thr}$ were showed stable canonical clover-leaf tRNA structures with acceptible anticodons. However, both the DHU and T$\psi$C arms of tRN $A^{Phe}$ could not form any stable stem-loop structure. The two overlapping gene pairs (tRN $A^{Lys}$ -tRN $A^{ASP}$ and ATPase8-ATPase6) were found from our sequencing results. The genes are encoded on the same strad. ATPase8 and ATPase6 overlaps (ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common, and there is evidence that the two proteins are transcribed from a single bicistronic message by initiation at 5$^1$terminal start site for ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mandarina) which is estimated as ancestal of domesticated silkworm.sticated silkworm.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.260-266
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• 2005

본 실험에서는 MRF가 다량 존재하는 RBL-2H3 세포주에 다양한 PKC isotype별 억제제를 처리하여 in vitro상에서 Na, K-ATPase $\alpha$1L3를 이용한 pull-down assay와 RBL-2H3 세포를 이용한 membrane fractionation을 실시하였다. 그 결과 HRF는 in vitro에서 $\alpha$1L3와 결합한다는 사실을 재확인 할 수 있었고 실제 세포주 내에서 Na,K-ATPase와 결합한다는 것을 알 수 있었다. 사용한 약물로부터 PKC $\alpha,\;\beta,\;\delta$뿐 아니라 protein phosphatase 2B(PP2B)도 HRF와 $\alpha$1L3의 결합에 관여한다는 사실을 알 수 있었다. 또한 이들 PKC, PP2B에 의해 인산화된 HRF 분자는 cytosolic fraction으로 이행할 수 있으며 이러한 결과는 탈인산화된 HRF가 Na,K-ATPase와 결합하여 Na, K-ATPase의 기능을 조절한다고 추정할 수 있다. 그러나 약물자체가 histamine 분비에 영향을 미칠 수 있으며 cytosolic HRF보다 exocytosis된 HRF가 histamine를 더 분비하도록 할 수 있으므로, 약물을 전처리한 세포에 외부에서 HRF를 첨가하여 histamine이 유리되는 정도가 어떻게 변화하는지를 HRF를 가하지 않은 결과와 비교해야 할 것이다.

• The Korean Journal of Physiology
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• 제8권1호
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• pp.55-65
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• 1974

The effect of ginseng on the ATPase activity of rabbit ref cell membrane has been investigated. The experiments were also designed to determine whether the components of ginseng could be attributed to the effect on ATPase activity which dependent upon sodium plus potassium and is sensitive to ouabain. The following results were observed. 1. The activity of the $Na^{+}-K^{+}-ATPase$ from red cell membrane is stimulated by ginseng, and the concentration of ginseng for half-maximal activity is about 15 mg%. The pH optimum for the ginseng sensitive component is 7.6. 2. The portion of the enzyme activity stimulated by ginseng is completely abolished by ouabain. 3. The activating effect of ginseng on the ATPase, with a given concentration of sodium in the medium, is increased by raising the potassium concentration but activity ratio is decreased. 4. The activating effect of ginseng on the ATPase, with a given concentration of potassium in the medium, is increased by raising the sodium concentration but the activity ratio is decreased. 5. The ATPase activity is increased by small amounts of calcium but inhibited by larger amounts and the rate of activity by ginseng is constant. 6. The action of ginseng on the ATPase activity was not related to the sulfhydryl group of cysteine, the amino group of lysine, the imidazole group of histidine, the quanidinium group of arginine, the carboxyl group of aspartic acid, or the hydroxyl group of threonine. 7. The activating effect of ginseng on the ATPase activity may be not due to a saponin which is contained in ginseng.

• 대한약리학회지
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• 제8권1호
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• pp.31-40
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• 1972

The author studied the actions of ouabain and diphenylhydantoin sodium on the ATPase activity in mitochondrial fraction isolated from rabbit heart and compared with that of quinidine. The results obtained are as follows: 1) In studying the $(Na^++K^+)-activated$ ATPase activity, the rabbit heart isolated was immediately frozen for 7-9 days (ageing of preparation) and thereafter the mitochondria1 fraction obtained by differential centrifugation technic was treated with solution A containing 0.15% deoxycholate for 24-48 hours at $-10^{\circ}C$ before using in experiment. These methods increased the activity ratio to 0.87-0.98. 2) The $(Na^++K^+)-activated$ ATPase activity in mitochondrial fraction of rabbit heart was not completely but markedly inhibited by ouabain. This inhibitory action of ouabain was moderately antagonised by $K^+$ concentration at constant Na concentration. 3) Diphenylhydantoin sodium in concentration of $5{\times}10^{-4}{\sim}10^{-3}M$ stimulated markedly not only $Mg^{++}-dependent$ ATPase activity but also $(Na^++K^+)$-activated ATPase activity and in concentration lower than $10^{-6}M$ had little effect. However, this effect of diphenylhydantoin was markedly increased in the presence of $Na^+$ alone rather than $K^+$ alone, but lesser than that effect in the presence of both $Na^+$ and $K^+$, together. The stimulating effect of diphenylhydantoin was specifically antagonized by ouabaion. 4) When the rabbits were intravenously injected with ouabain and diphenylhydantion respectively, $(Na^++K^+)-activated$ ATPase activity of rabbit heart of ouabain-treated group was much decreased and both $(Na^++K^+)-activated$ ATPase and $Mg^{++}-activated$ ATPase activity were moderately increased in diphenylhydantoin-treated rabbit group. 5) The $(Na^++K^+)-activated$ ATPase activity in mitochondrial fraction of rabbit heart was slightly inhibited by quinidine in high concentration of $10^{-4}M$, but nearly little effect was observed below the concentration of $5{\times}10^{-5}M$. 6) It might be possible to conclude that diphenylhydantoin specifically antagonised the action of ouabain on the membrane ATPase, which is different from the action of quinidine.

• 한국식품영양과학회지
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• 제19권4호
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• pp.349-355
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• 1990

Actomyosine prepared from Yellowtail fish(seriola quinqueradiata) were stored at $0^{\circ}C$(ice-cooling) -3.5$^{\circ}C$(partial freeaing) and -2$0^{\circ}C$(freezing) Another actomyosin samples were prepared from the fish previously stored at the temperatures for a week as the maximum .Remaining activity of {{{{ {Ca }^{2+ } }}}}-and {{{{ {Mg }^{2+ } }}}}- dependent adenosine triphosphatase(ATPase) activity was measured fronm the actomyosin preparations. Specific activity of {{{{ {Mg }^{2+ } }}}}-ATPase of actomy-osin before storagew was 0.253$\mu$ mole pi/min/mg of protein and it was 1.5 times higher than that of {{{{ {Ca }^{2+ } }}}} -ATPase. The enzyme activities were markedly decreased during early period of storage. However no significant differences in the enzyme activity were revealed among the samples stored at different temperature. The enzyme of actomyosin prepared from the fish previously stored at the temperatures for a week revealed an acitivity of 2-3 times higher than that of freezing. Apparent denaturation constant of {{{{ {Mg }^{2+ } }}}} -ATPase of actomyosin was between 0.810-1.139 per day and it was about 1.5 times hgiher than that of {{{{ {Ca }^{2+ } }}}} -ATPase. But the constant of {{{{ {Mg }^{2+ } }}}} ATPase of actomyosin extracted from the fist stored for a week at each temperature was between 0.176-0.356 per day. This constant was 4 times higher than that of {{{{ {Ca }^{2+ } }}}}- ATPase in frozen stored fish. It was presumed from these results that denaturation of ATPase is largely accorded to the structural changes of actomyosin.